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Cyanobacterial Photosystem II at 3.2 Å resolution – the plastoquinone binding pockets

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Abstract

Photosystem II from thylakoid membranes of the thermophilic cyanobacterium Thermosynechococcus elongatus was solubilized with n-β-dodecylmaltoside and purified using anion exchange chromatography. Molecular weight, pigment stoichiometry and subunit composition were assayed using various techniques. The holocomplex is dimeric with a molecular mass of 756 ± 18 kDa and functionally fully active. Crystals obtained from these samples showed significantly improved quality leading to a 3D structure at 3.2 Å resolution. Several loop regions of the principal protein subunits are now defined that were not interpretable at lower (3.8 Å) resolution, thus resulting in a more complete model. The head groups of the cofactors of the electron transfer chain and of the antennae have been modeled, coordinating and hydrogen bonding amino acids identified and the nature of the binding pockets derived. The orientations of these cofactors resemble those of the reaction centre from anoxygenic purple bacteria. For the two plastoquinones, electron density was only found for the head group of Q A and none for Q B indicating low or even no occupancy of this site in the crystal structure. Both binding pockets and problems related to the Q B site are discussed here and compared to the situation in the purple bacterial reaction centre.

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Abbreviations

AAS:

atom absorption spectroscopy

β-DM:

n-β-D-dodecylmaltoside

Car:

β-carotene

Chl a:

Chlorophyll a

cyt:

cytochrome

DLS:

dynamic light scattering

GPC:

gel permeation chromatography

MALDI-TOF MS:

matrix assisted laser desorption ionisation time of flight mass spetrometry

PbRC:

purple bacterial reaction centre

Pheoa:

Pheophytin a

PQ9:

plastoquinone 9

PS:

photosystem

Q A :

primary quinone

Q B :

secondary quinone

RPC:

reverse phase chromatography

TLC:

thin layer chromatography

UC:

ultracentrifugation

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Authors and Affiliations

  1. Max-Volmer-Laboratories for Biophysical Chemistry, Technical University Berlin, Strasse des 17 Juni 135, 10623, Berlin, Germany

    Jan Kern, Athina Zouni & Klaus-Dieter Irrgang

  2. Institute of Chemistry/Crystallography, Free University Berlin, Takustrasse 6, 14195, Berlin, Germany

    Bernhard Loll, Wolfram Saenger & Jacek Biesiadka

Authors
  1. Jan Kern
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  2. Bernhard Loll
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  3. Athina Zouni
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  4. Wolfram Saenger
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  5. Klaus-Dieter Irrgang
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  6. Jacek Biesiadka
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Corresponding authors

Correspondence to Klaus-Dieter Irrgang or Jacek Biesiadka.

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Kern, J., Loll, B., Zouni, A. et al. Cyanobacterial Photosystem II at 3.2 Å resolution – the plastoquinone binding pockets. Photosynth Res 84, 153–159 (2005). https://doi.org/10.1007/s11120-004-7077-x

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  • DOI: https://doi.org/10.1007/s11120-004-7077-x

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